Cioni, Carla

D'Este, Loredana

Parisi Salvi, Ebe

Toni, Mattia

Vivacqua, Giorgio

Yu, Shun

Firenze University Press: E-Journals

IJAE Vol. 115, n. 1/2 (Supplement), 2010
178
Identification of α-synuclein in the CNS of lower 
vertebrates by two novel monoclonal antibodies: first 
immunohistochemical and western blot evidence
Giorgio Vivacqua1, Ebe Parisi Salvi1, Carla Cioni2, Mattia Toni2, Shun Yu3, Loredana D’Este1
1 Department of Human Anatomy, Faculty of Medicine, Sapienza University of Rome, Italy
2  Department of Animal and Human Biology, Faculty of Sciences, Sapienza University of Rome, Italy
3 Department of Neurobiology, Xuanwu Hospital and Capital Medical University, Beijing, China
Α-Synuclein (α-Syn) is a 140 aa protein which is highly expressed in the CNS of dif-
ferent vertebrates and linked to Parkinson’s disease (PD) in mammals. We have exam-
ined the localization of α-Syn in the CNS of zebrafish (Danio rerio), a widely used model 
species for PD, the carp (Cyprinus carpio) and the African clawed frog (Xenopus laevis) 
through immunohistochemistry and western blot (WB) by using two new monoclonal 
antibodies (2E3 and 3D5) against α-Syn. In mammals, both antibodies labeled nerve 
fibers and 3D5 also localized α-Syn within the neuronal nuclei. Conversely to mam-
mals, no nuclear staining was detectable in fish and amphibian neuronal bodies by 3D5. 
Moreover, the anatomical distribution of the protein is markedly different. In fact, in 
mammals α-Syn is ubiquitary distributed in the brain and the spinal cord, whereas in 
lower vertebrates only few brain regions display a positive staining. In accordance with 
literature data, these regions provide the amount of dopamine and catecholamines to 
the brain. Therefore, our results suggest that in lower vertebrates α-Syn is exclusively 
linked to catecholaminergic and dopaminergic systems and it may represent a protein 
linked to synaptic vesicles, whereas in mammals it becomes an invariant and structural 
component of distinct neuronal subtypes assuming a new subcellular localization. WB 
analysis performed on the CNS homogenates by 2E3 antibody revealed an intense band 
at 19 kDa. However, the 3D5 antibody stained only faint bands in the same homoge-
nates. These results confirm the presence of α-Syn in the CNS of lower vertebrates ana-
lyzed. Our observation suggest that the role of α-Syn in the neuronal physiology could 
improve, during the evolution of vertebrates and phylogenetic approach could repre-
sent a novel method to understand the physio-pahological role of α-Syn in neurological 
diseases.
Key words
Alpha-Synuclein, Danio rerio, Xenopus laevis, Immunohistochemistry, Western blot, Parkinson 
disease


Identification of α-synuclein in the CNS of lower vertebrates by two novel monoclonal antibodies: first immunohistochemical and western blot evidence

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Identification of α-synuclein in the CNS of lower vertebrates by two novel monoclonal antibodies: first immunohistochemical and western blot evidence

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